pBSep1 Vector

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Testing B.subtilis promoters and RBSs

We are seeking to test the expression strength and response to inducers in several promoters in B.subtilis, in combination with different Bacillus-specific ribosome binding sites.

B. subtilis Promoter Testing with Beta-galactosidase Assay

We are aiming to characterize four different Bacillus subtilis promoters using the beta-galactosidase assay. (Protocol for beta-galactosidase assay as described here

B. subtilis Promoters to be Tested:

• Pupp
• Ppac
• Pspac
• Pxyl

Links

• Chris French's protocol on transforming B. subtilis
• We have last year's protocol; not sure if it's trustworthy, need to compare with online resources
• Protocol from BCSG catalog
  • used in Cornell bacteriology course, probably should start here

B. Subtilis Transformation Protcol - Electroporation

Bongers et al. … of a Subtilin-Regulated Expression System in Bacillus subtilis: Strict Control of Gene Expression …. Applied and Environmental Microbiology (2005)

• SURE: handy expression system for B. subtilis
• What about feedback/regulation effects of subtilin on the cell

Expression and characterization of aiiA gene from Bacillus subtilis BS-1.

• (Some strains of) Bacillus subtilis produces a gene called aiiA that degrades AHL molecules from gram-negative bacteria
• this is biobricked

Links to Cambridge 2007 Wiki

• B. subtilis group
• B. subtilis group: more info